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Here, a previously uncharacterized CE7 family acetylxylan esterase (TsAxE) from Thermoclostridium stercorarium was identified and engineered through a parallel multidimensional engineering strategy integrating consensus motif engineering, molecular docking, and interface engineering. The best-performing mutant BCF (D218L\/D245P\/G183Y) exhibited a 44.10 \u00b1 0.44% increase in activity toward p-nitrophenyl acetate (pNPA). Notably, the thermostability of BCF was substantially improved, with the half-life (t1\/2) at 60 \u00b0C extended from 0.63 \u00b1 0.04 h to 43.82 \u00b1 3.90 h. In addition, BCF showed improved catalytic efficiency toward p-nitrophenyl butyrate (pNPB). Molecular docking and molecular dynamics analyses suggested that these mutations may reshape substrate-binding pocket and improve structural stability. This study demonstrates the effectiveness of integrating complementary engineering strategies for the rational improvement of AxEs activity and thermostability.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c04818","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T04:02:29Z","timestamp":1785902549000},"page":"25436-25451","source":"Crossref","is-referenced-by-count":0,"title":["Parallel Multidimensional\nEngineering of Acetylxylan\nEsterase Enables Concurrent Enhancement of Thermostability and Catalytic\nEfficiency"],"prefix":"10.1021","volume":"74","author":[{"given":"Yujiao","family":"Tao","sequence":"first","affiliation":[{"name":"Jiangsu University , , 301 Xuefu Road , ,","place":["Zhenjiang, China, 212013"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinrui","family":"Tang","sequence":"additional","affiliation":[{"name":"Jiangsu University , , 301 Xuefu Road , ,","place":["Zhenjiang, China, 212013"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-2241-4813","authenticated-orcid":true,"given":"Mei","family":"Zhao","sequence":"additional","affiliation":[{"name":"Jiangsu University , , 301 Xuefu Road , ,","place":["Zhenjiang, China, 212013"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjing","family":"Sun","sequence":"additional","affiliation":[{"name":"Jiangsu University , , 301 Xuefu Road , ,","place":["Zhenjiang, China, 212013"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5817-1609","authenticated-orcid":true,"given":"Xianghui","family":"Qi","sequence":"additional","affiliation":[{"name":"Jiangsu University , , 301 Xuefu Road , ,","place":["Zhenjiang, China, 212013"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,4]]},"reference":[{"key":"2026081910174189600_cit1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcsw.2023.100101","article-title":"An update on xylan structure, biosynthesis,\nand potential commercial applications","volume":"9","author":"Curry","year":"2023","journal-title":"Cell\nSurf."},{"issue":"7","key":"2026081910174189600_cit2","doi-asserted-by":"publisher","DOI":"10.3390\/fermentation9070606","article-title":"Discovery of a novel \u03b2-xylosidase with xylanase\nactivity and its application in the production of xylitol from corncob\nxylan","volume":"9","author":"Liang","year":"2023","journal-title":"Fermentation"},{"issue":"7","key":"2026081910174189600_cit3","doi-asserted-by":"publisher","first-page":"1260","DOI":"10.1016\/j.biotechadv.2016.09.001","article-title":"Towards enzymatic\nbreakdown of complex\nplant xylan structures: State of the art","volume":"34","author":"Biely","year":"2016","journal-title":"Biotechnol.\nAdv."},{"key":"2026081910174189600_cit4","doi-asserted-by":"publisher","DOI":"10.1016\/j.carbpol.2022.119271","article-title":"Gut microbial\nutilization of xylan and its implication in gut homeostasis and metabolic\nresponse","volume":"286","author":"Zhang","year":"2022","journal-title":"Carbohydr. 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